摘要
为探究CBP酿酒酵母菌群表面展示嵌合纤维小体的协同作用,该文通过组装纤维素酶和半纤维素酶进行木质纤维素乙醇发酵的性能,构建由分别可表面展示纤维素酶嵌合纤维小体(由支架蛋白ScafI,内切葡聚糖酶、外切葡聚糖酶和β-葡糖苷酶构成)和半纤维素酶嵌合纤维小体(由支架蛋白ScafII,内切木聚糖酶和外切木糖苷酶构成)的重组酿酒酵母菌EBY100株组成的CBP菌群。通过蛋白质的SDS-PAGE、Western Bolt验证、酶活性测定、细胞表面定位和纤维小体组装效率分析,证明了融合蛋白能够以活性形式组装成2种形式的嵌合纤维小体。以蒸汽爆破玉米秸秆为底物进行同步糖化发酵产乙醇,96 h时最大乙醇产量达到0.673 g/L,消耗每克碳水化合物的乙醇产量0.255 g,相当于乙醇产率理论值的50.1%,协同因子为1.06。结果证明两株酵母细胞表面的不同类型嵌合纤维小体在水解木质纤维素过程中存在明显的协同促进作用。
In order to investigation the synergistic effect of cellulases and xylanases displaying on CBP yeast cell surface,the cellulasecellulosome consisting of endoglucanase,exoglucanase and β-glucosidase,and the hemicellulase cellulosome consisting of xylanaseand xylosidase was self-surface assembly and investigated. SDS-PAGE,Western,enzyme activity,immunofluorescence micrographs and flow cytometry analysis proved that two types of designer cellulosome were assembled. The final ethanol concentration from steamexploded corn stalk was up to 0.673 g/L 96 h after fermentation and the overall yield was 0.255 g of ethanol produced per gram of carbohydrate consumed,corresponding to 50.1% of theoretical value,and the degree of synergy was up to 1.06 finally. These results confirmed the synergistic effect of cellulase and xylanase binded to the designer cellulosome which anchored on the two yeast strains respectively.
作者
杜济良
刘磊
万平
杨秀山
田沈
Du Jiliang;Liu Lei;Wan Ping;Yang Xiushan;Tian Shen(College of Life Science,Capital Normal University,Beijing 100048,China)
出处
《太阳能学报》
EI
CAS
CSCD
北大核心
2021年第1期342-347,共6页
Acta Energiae Solaris Sinica
基金
国家科技支撑计划(2019YFB1503802)
国家自然科学基金(31971202)
科技创新服务能力建设资助项目(025185305000/198)。
关键词
酿酒酵母
纤维素乙醇
生物质
细胞表面展示
嵌合纤维小体
统合生物加工
Saccharomyces cerevisiae
cellulosic ethanol
biomass
cell surface display
designer cellulosome
consolidated bioprocessing
